Comparative studies on similarities and differences of cyclodipeptide oxidases for installation of C–C double bonds at the diketopiperazine ring
| Autor: | Shu-Ming Li, Rixa Kraut, Johanna Schäfer, Lena Mikulski, Kirsten Brockmeyer |
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| Rok vydání: | 2020 |
| Předmět: |
Double bond
Stereochemistry Diketopiperazines Ring (chemistry) Methylation 01 natural sciences Applied Microbiology and Biotechnology Catalysis Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Prenylation Escherichia coli Aromatic amino acids Amino Acids Biotechnologically Relevant Enzymes and Proteins Biotransformation 030304 developmental biology chemistry.chemical_classification 0303 health sciences Cyclodipeptides and derivatives biology 010405 organic chemistry General Medicine Carbon Enzyme assay 0104 chemical sciences Amino acid Enzyme chemistry biology.protein Cyclodipeptide oxidase Dehydrogenation Oxidoreductases Biotechnology |
| Zdroj: | Applied Microbiology and Biotechnology |
| ISSN: | 1432-0614 0175-7598 |
| DOI: | 10.1007/s00253-020-10392-7 |
| Popis: | Abstract Cyclodipeptide oxidases (CDOs) perform dehydrogenations on diketopiperazines and play an important role in the cyclodipeptide diversification. In this study, we investigated the two known CDOs AlbA/B and Ndas_1146/7 and one new member, CDO-Np. LC-MS monitoring of 32 cyclodipeptide biotransformations in E. coli revealed good consumption of cyclodipeptides containing aromatic amino acids. Cyclodipeptides consisting solely of aliphatic amino acids were poor substrates. In vitro assays of 34 substrates with crude enzyme extracts and product identification proved that the CDO-Np-containing extract catalyzes the formation of two C–C double bonds in many cases. The extracts containing the two other enzymes had lower activities and catalyzed mainly didehydrogenations. For didehydrogenation, the phenylalanyl or tyrosyl site was usually preferred. No or very low acceptance of benzodiazepinediones and a 2,6-diketopiperazine proved the importance of the 2,5-diketopiperazine ring. N-Methylation at the diketopiperazine ring or prenylation of the tryptophan-containing cyclodipeptides influences the enzyme activity and product spectrum. Key points • Comparison of catalytic activities of three enzymes; Diverse cyclodipeptides and derivatives as substrates; Determination of double bond formation using2H-labeled substrates; Product identification also by interpretation of MS2fragmentation pattern. |
| Databáze: | OpenAIRE |
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